A novel emulsion mixture for in vitro compartmentalization of transcription and translation in the rabbit reticulocyte system
Open Access
- 1 March 2004
- journal article
- research article
- Published by Oxford University Press (OUP) in Protein Engineering, Design and Selection
- Vol. 17 (3) , 201-204
- https://doi.org/10.1093/protein/gzh025
Abstract
Emulsion formulations used for in vitro compartmentalization (IVC) methods were found to be incompatible with protein expression in the rabbit reticulocyte (RRL) system, causing rapid discoloration and translation shutdown. Here we identify possible causes and describe a novel water‐in‐oil emulsion which abolished discoloration and allowed high‐level in‐emulsion expression of active luciferase and human telomerase using the RRL. This novel emulsion greatly expands the range of potential protein targets for IVC.Keywords
This publication has 26 references indexed in Scilit:
- Altering the sequence specificity of HaeIII methyltransferase by directed evolution using in vitro compartmentalizationProtein Engineering, Design and Selection, 2004
- Directed evolution of an extremely fast phosphotriesterase by in vitro compartmentalizationThe EMBO Journal, 2003
- Strategies for selection of antibodies by phage displayPublished by Elsevier ,2002
- In vitro display technologies: novel developments and applicationsCurrent Opinion in Biotechnology, 2001
- Directed evolution of polymerase function by compartmentalized self-replicationProceedings of the National Academy of Sciences, 2001
- Translation initiation: adept at adaptingTrends in Biochemical Sciences, 1999
- STABLE: protein‐DNA fusion system for screening of combinatorial protein libraries in vitroFEBS Letters, 1999
- Antibody-ribosome-mRNA (ARM) complexes as efficient selection particles for in vitro display and evolution of antibody combining sitesNucleic Acids Research, 1997
- In Vitro Translation and Assembly of a Complete T Cell Receptor–CD3 ComplexThe Journal of Experimental Medicine, 1997
- Cell-free synthesis and assembly of connexins into functional gap junction membrane channelsThe EMBO Journal, 1997